title: Construction of a plasmid for co-expression of mouse membrane-bound form of IL-15 and RAE-1ε and its biological activity.
abstract: Interleukin 15 (IL-15) is a pivotal cytokine for the proliferation and activation of a specific group of immune cells such as natural killer (NK), IFN-producing killer dendritic cells (IKDC) and CD8 T cells. RAE-1ε, the ligand for the activating NKG2D receptor, which also play an important role in the proliferation and activation of NK cells and IKDCs. In this study, a membrane-bound form of IL-15 (termed mb15) encoding sequence and RAE-1ε gene were obtained by SOE-PCR or PCR amplification. The amplified mb15 and RAE-1ε gene were then digested and inserted into the multiple cloning site1 (MCS1) and MCS2 of pVITRO2-mcs vector, respectively. A recombinant eukaryotic expression vector for co-expression of mb15 and RAE-1ε was successfully constructed. After it was transfected to BaF3 cells, the expression of IL-15 and RAE-1ε in recombinant BaF3/mb15/RAE-1ε cells were verified by RT-PCR, western blot and FCM analysis. Furthermore, BaF3/mb15/RAE-1ε cells had the ability of promoting NK cells proliferation and IFN-γ secretion. In conclusion, BaF3/mb15/RAE-1ε cells were successfully constructed, which is very useful for further studies, especially for the expansion and activation of certain subsets of immune cells such as NK cells and IKDCs.
authors.affiliation: Department of Immunology, School of Medicine, Yangzhou University, Yangzhou 225001, PR China. liqianxh@163.com
authors.forename: Li
authors.initials: L
authors.lastname: Qian
authors.forename: Ming-Chun
authors.initials: MC
authors.lastname: Ji
authors.forename: Xin-Yuan
authors.initials: XY
authors.lastname: Pan
authors.forename: Wei-Juan
authors.initials: WJ
authors.lastname: Gong
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
authors.forename: Qiu-Fang
authors.initials: QF
authors.lastname: Duan
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D015870:Gene Expression / Q000235:genetics*; D005786:Gene Expression Regulation / Q000276:immunology; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D019409:Interleukin-15 / Q000235:genetics* / Q000378:metabolism*; D007694:Killer Cells, Natural / Q000276:immunology / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D055655:NK Cell Lectin-Like Receptor Subfamily K / Q000378:metabolism; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism*
chemical_list: D019409:Interleukin-15; D008565:Membrane Proteins; D055655:NK Cell Lectin-Like Receptor Subfamily K